Method of preparing zein solutions directly from gluten



Gluten.

Patented May 29, 1945 METHOD OF PREPARING ZEIN SOLUTIONS DIRECTLY FROM GLUTEN Roy E.

Coleman. Chicago, 111., assignor to Time,

Incorporated, a corporation oi New York No Drawing. Original application November 30,

1942, Serial 2,352,804, dated No. 467,402, new Patent No. July 4, 1944. Dividedandthis application October 25, 1943, Serial No. 507,598

8 Claims. (Cl. 106-453) This invention relates to methods oi preparing commercially usable solutions and coating compositions oi prolamines or prolamine-containing proteins directly irom glutens, either corn, wheat, barley, etc. It is particularly directed to such methods which will produce solutions and coating compositions of the alcohol-soluble portion of corn gluten, oi controlled solution and gelling characteristics.

This application is a division of my copending application, Serial No. 467,402, filed November 80, 1042, now Patent No. 2,352,604, dated July 4, 1044.

Beretoiore. in the preparation oi zein solutions and coating compositions, including the substantially non-aqueous zein solutions and coating compositions of controlled solution and gelling characteristics as described in my prior Patent No. 2,185,110, granted December 26, 1939, for example, the zein is initially isolated from corn gluten and then dissolved in suitable solvents to form the desired solutions. To isolate zein irom corn gluten as a commercially usable powder is a laborious and expensive process, involving. steps of precipitation. settling, filtering. washing and drying. The solubility of the zein in zein solvents and the solution characteristics such as stability and gelling tendencies are largely dependent upon the care exercised in carrying out the steps of the process.

In accordance with the present invention I prepare solutions or zein or zein-containing proteinl directly from corn gluten under conditions that will readilyproduce a wide variety of zein products in so far as properties and solubility characteristics are concerned, and thereby avoid theexpense and objections incident to the preliminary isolation of zein in powder form. In carrying out the present invention, corn gluten, ior example; in granular or powdered form. is treated with a suitable solvent mixture, without the establishment of special conditions of hydrogen ion concentration either in the solvent or in the gluten, to extract the zein from the The solvent mixture comprises a base solvent having a boiling point above about 125 0., preferably above about 150 C. and a diluent or extender having a boiling point below about 125 C. and, preferably, of about C. and below. The base solvent is itself a solvent for coin and the diluent may be, and preferably is, also a solvent to: rein The base solvents and diluents in with the present invention will be defined fully hereafter.

The extraction is carried out at elevated temperatures, say from about to about 170 F., preferably between about to about F., 101' a short period oi time generally not exceeding about three hours. It is preferred that the extraction time be not longer than about One-half to about one hour since within this time and at the temperatures stated, the rein in the gluten is extracted and is not deleteriously afiected.

The extract solution obtained as above described is now separated from the residual gluten by filtration, pressing. centrifugal separation or in any other desirable manner and is distilled or otherwise treated to remove all or some of the diluent, as desired. It may be filtered preliminarily to the step of removing the diluent, if necessary. The resulting residue is a solution of zein or zein-containing protein in the base solvent alone with all of the diluent irom the extract solution removed. or in the base solvent and that portion 01' the diluent which has not been removed. The residual gluten, which still contains some protein, may be freed from retained solvent by washing and recovered for further use as a cattle loud or for other purposes.

The resulting zein or zein-containing solutions in accordance with the present invention, over a wide range of concentrations, are stable on standing at temperatures of 80 F. and below and do not separate even when cooled to temperatures oi 50 to 70 F. and somewhat below. When cooled even to temperatures as low as 10 above zero or to zero. solutions prepared in accordance with the present invention may separate and become solid: however, on from about 45 to 70 F., a resolution is effected, either without stirring or with slight stirring, and the solutions return to their normal form at the reheating temperatures. In general, these solutions are substantially non-gelling and have but a slight tendency to increase in viscosity; that is, to thicken or become heavier in body. In many instances they exhibit practically no tendency to thicken or to gel over practical periods of time in the order 0! about 3 to 6 months and even up to one year and longer. These solutions remain stable and reasonably constant in their characteristics. during the periods necessary for transmutation and distribution, either in bulk or in packages, or for storage for reasonable periods.

0! time, and hence their field of applicability is greatly increased.

In general, in the concentration of zein or other prolamine in commercially usable solutions containing about 12 to 30% oi protein, the

amount of base solvent necessary to obtain these heating again to temperatures or concentrations in the final solutions is in and of itself insufficient to give the proper relation between solvent and gluten for eifective extraction results. If large amounts of the relatively high boiling point base solvents are used, sumcient to give the proper relation between solvent and gluten, the extraction results are not suillciently eflective and, moreover, the cost of treating the extracts to obtain the desired solids concentration is so prohibitive as to make this procedure commercially unfeasible.

The desired solvent to gluten ration in accord- 1 ance with the present invention is secured by adding to the desired amount of relatively high boiling point base solvent or mixture of base solvents one or more of the so-called relatively low boiling point diluent solvents. To obtain eiiective extraction results, the solvent to gluten ratio should be in the order of about 2 to parts by weight of the former to about 1 part by weight of the latter. Due to the presence of a diluent or extender in the extracting solvent, the solvent to gluten ratio may be less than that which would be required with abase solvent alone and, notwithstanding the lower solvent to gluten ratio, the extraction results are generally more effective. The diluent solvent or extender must have a lower boiling point and distillation range than the base solvent. It is preferred that these diluent solvents or extenders be of substantially lower boiling point and distillation range than the base solvent, as pointed out above.

The base solvents are relatively high boiling compounds or mixtures of compounds as pointed out above and have a suitable balance between the hydrocarbon constituents or radicals present in the solvent or solvent mixtures and the polar constituents or radicals present therein. (A certain proportion of hydroxyl radicals in the base solvents is required to secure solvent action. Other polar radicals such as O (ether oxygen), Cl, NH: and COOH appear to aid or cooperate with the OH in securing the necessary conditions of polarity in the base solvents (in which term I include mixtures of base solvents), for solvent action.

The proportion of polar radicals to hydrocarbon or non-polar radicals which I have found to be desirable lies within a range which extends between the limits of absolute methanol on the one hand, having 53% hydroxyl and 47% hydrocarbon radicals, and absolute ethanol on the other hand having 37% hydroxyl and 63% hydrocarbon radicals. The benzyl radical acts as if it were intermediate the methyl and ethyl radicals. This range is approximate and may vary somewhat depending on the solvent or solvent mlxture used. The limits of this range are indicative of solvent properties and provide a close and adequate guide to enable the operator, by simple experimentation, to determine the required balance of polar and hydrocarbon radicals in the base solvent or mixture of base solvents in accordance with the present invention.

It may be stated that the base solvents in accordance with the present invention may be any one or the solvents or mixture of solvents described ln my prior Patent No. 2,185,110 which has the required balance of polar and non-polar radicals, and a boiling point above about 125 0., preferably, above about 150 C. as pointed out above. For a more complete description of the that of solvents which may be used as base ear-1,290

solvents in accordance with the present invention than is disclosed herein. reference is made' to the descriptionof these solvents in my prior Patent No. 2,185,110.

Examples of base solvents in accordance with the present invention are the glycols such as diethylene glycol, propylene glycol, triethylene glycol and mixtures thereof, including mixtures which contain ethylene glycol; diacetone alcohol; closed chain cyclic alcohols such as benzyl alcohol, cyclohexanol, furfuryl alcohol, tetrahydrofuriuryl alcohol and mixtures of closed chain alcohols; glycol ethers such as ethylene glycol monomethyl ether, diethylene glycol monomethyl ether, ethylene glycol mono-ethyl ether, diethylene glycol monoethyl and mixtures of glycol ethers; mixtures of two or more of the foregoing enumerated base solvents; mixtures of one or more of the foregoing base solvents with glycerine, or other solvents or mixtures of solvents, providing the mixture has the required polar to non-polar radical balance and the required relatively high boiling point. It is to be understood that the foregoing specifically mentioned solvents or mixtures of solvents is not intended to be inclusive of all the base solvents which may be used in accordance with the present invention since they are merely set forth for illustrative purposes.

Diluent solvents mixtures (azeotropes) of aliphatic alcohols such as 96% ethyl alcohol and 91% isopropyl alcohol. For convenience herein I term these alcoholic solvents "concentrated alcohol solvents." The diluent solvents may also be one or more of the concentrated alcohol solvents containing additional amounts of water, say from 40% to 50% in anhydrous methanol or in excess of that'present in the azeotropes alcohol and 81% isopropyl alcohol. Greater or lesser amounts of water than as stated may be used; however, best results are obtained with the use of added amounts of water within the percentage range stated. For convenience herein I term these alcoholic solvents "aqueous alcohol mixtures.

Another class of diluent solvents in accordance with the present invention comprisesmixtures of an azeotropic concentrated alcohol solvent with a non-solvent and water. Examples of such diluents are mixtures of 95% ethyl alcohol or 91% lsopropyl alcohol with dichlorethylene, trichlorethylene, ethyl acetate, benzene, cyclohexane,

etc., and water. In this class 0! diluent solvents I also include non-aqueous azeotropic mixtures of anhydrous methyl alcohol with hexane; cyclohexane, benzene, carbon tetrachloride, dichlorethylene, trichlorethyiene, acetone, methyl acetate, ethylacetate, methyl ethyl ketone, etc.. because one constituent of the mixture. methyl alcohol, is a zein solvent.

In addition to the foregoing, wherein at least one constituent of a diluent solvent mixture is a zein solvent, the diluent solvents may be suitable azeotropic mixtures which are zein protein 'solvents wherein the individual constituents of the mixture are non-solvents tor sein proteins. Thus. they may be as eotropic mixtures of water on the one hand and dioxan or butyl alcohol on the other. Dioxan forms with water a constant boiling point mixture containing 80% dioxan boiling at 88.9 (3.; and butyl alcohol forms with water a constant boiling point mixture containing 63% butyl alcohol boiling at 92 c. They may also be suitable aqueous aseotropic mixtures which are non-solvents for zein proteins such as ethyl methyl ketone and water. also be non-aqueous azeotropic mixtures of anhydrous ethyl alcohol or anhydrous isopropyl alcohol with hexane, cyclohexane, benzene. carbon tetrachloride, dichlorethylene. trichlorethylene. acetone, methyl acetate, ethyl acetate, methyl ethyl ketone, etc.

'I'he diluents may also be suitable non-solvents or low polarity characterizedby the presence of -O. --Cl, NH:, -COOH groups, etc. Examples of such diluents are dioxan, dichlorpentane, ethylene dichloride, dichlorethyl ether. etc. They may also be miscible hydrocarbons such as bensenahydrogenated naphtha. etc. Or they may be suitable non-solvents of high polarity And they may such as anhydrous ethanol, anhydrous isopro traction procedures, using either a concentrated alcohol solvent alone or an aqueous alcohol mixture alone, approximately 2 parts by weight oi solvent to 1 of gluten may be used. At this concentration, with gluten containing approximately 60% total protein, the extract solutions of the concentrated alcohol solvent contain a maximum zein protein content of approximately 10% whereas the extract solutions of the aqueous alcohol mixture solvents contain a maximum zein protein content oi from about 12% to about 15% and even up to In addition, the aqueous alcohol mixture solvents are capable of extracting a substantially larger proportion of a desirable type of zein present in the protein oi the gluten than are the concentrated alcohol solvents.

Zein solutions of a, desired concentration and or a wide applicability can be obtained directly from gluten by having in the final solution as the only solvent os as the-main solvent for zein, a relatively high boiling point solvent having the required balance oi polar and non-polar redicals as described herein. It is now well established that the viscosity, gelling tendencies, and practical usability oi zein solutions are in no small part dependent upon the character oi the solvent in which the zein is dissolved.

Zein or zein-containing extract solutions containing any one o more or the foregoing base solvents in ECCOI'flflllCB with the present invention, may be boiled, distilled or otherwise treated to remove part or all of the diluent. It is evident that practically all of the diluent. including any water carried from the gluten to the extract solution, may be removed, thereby obtaining solutions of zein or zein-containing proteins, in the base solvent which, for all practical purposes, are substantially anhydrous. Such solutions vary widel in character, depending upon the choice of solvents or solvent mixtures used, as is to be expected, and some of them may undergo lined.

changes (apparent insolubillty, for example), during the diluent and water removal which atiect the solvent characteristics of the seln and zein-containing proteins in the base solvent uti- These' changes, where they take place, are not of a permanent character and it is possible to eil'ect a re-solution oi the main or seincontaining proteins in the base solvents by the addition of a small amount of water, say in the order of irom about 1 to 8% by weight, or an equivalent amount of other highly hydroxylated compound, such as methyl or ethyl alcohol or a mixture of such compounds.

In the preparation 01 usable solutions having a wide ileld of applicability, I prefer that the removal of diluent and water be controlled so as tial constituent of some of the solutions formed v in accordance with the present invention, the solutions are generally of an improved character when they contain small amounts of water. The solutions may contain larger amounts of water, say between 5 to 10%, or even more.

As already pointed out, the type or character oi the zein-containing protein produced by the extraction is in no small part dependent on the character of the extraction solvents used. This condition is particularly pronounced when the highly aqueous alcohols or the high water azeo- -tropes such as dioxan-water or n-butyl alcoholwater mixtures are used as diluents. Not only do these diluents elect the extraction of a larger amount of the zein-containing protein than do the more concentrated alcohols, but they also produce by their extraction a type of zein protein which generally produces solutions having a high viscosity and pronounced colloidal consistency. On the other hand the zelng protein produced with the concentrated alcohols and less aqueous diluents tends to produce solutions having a lower viscosity and a lesser colloidal consistency. It is thus evident that various types of zein protein having substantially diil'erent properties can be produced by proper selection and composition of the diluents.

From the foregoing it is manifest that the nature and properties of the finished solution will be determined by the nature of the base solvent, the amount of zein protein present in the solution and the type or character of the zein protein extracted by the particular diluent used. It is apparent that with the solvent and diluent combinations possible, a wide variety of solutions for many uses can be produced by the methods of the present invention.

Within the range of the many possible combinations and the methods in accordance with the present invention, solutions may be produced which require further adjustment or modifica tion in order to secure improved solution or desired stability and controlled gelling properties. Such adjustments or modifications can be readily effected by the use oi rosin, fatty acids, compatible amines such as monoethanolamlne, triethanolamine, etc., amine soaps of rosin or fatty acids, or by the various methods and materials for producing stable, controlled gelling, zein solutions described in my issued Patents Nos. 2,185,- 122, 2,246,779, and 2,298,548. The addition of suitable materials for shooting desired modification or adjustment may be made to the extract solution berore distilling 08 the diluent or to the final solution. The preferable procedure and materials with particular solutions may be easily determined by experimentation.

In the following examples I disclose illustrative methods for producing coin or zein-containing solutions directly from gluten in accordance with the present invention. In the examples and elsewhere throughout the specification. the term "parts" indicates parts by weight.

Example 1.-A solvent mixture containing 72 parts of anhydrous isopropyl alcohol (09%), 40 parts of diethylene glycol and 8 parts of ethylene glycol was added to 30 parts of powdered luten in a vessel. The mixture was heated to about 160 F. and held at about that temperature for about ,5 hour with stirring to extract the zein-containing proteins from the gluten. The mixture was then forced through a filter to separate the rein-containing extract and the extract was then slowly heated to about 250 F. and held at about that temperature with stirring until cessation of visible boiling. The residue was a solution of zein-containing proteins principally in the mixture oi glycols.

In the following examples the manipulative procedures were the same as those set forth in Example 1, with the exceptions hereinafter noted, and in each instance the extracts were heated until cessation of visible boiling.

Example 2.-A solvent mixture containing 72 parts of 91% isopropanol and 48 parts of di ethylene glycol monoethyl ether was added to- 36 parts of powdered gluten. The procedure was as in Example 1 except that the extraction temperature was 180' F. The resulting residue was a solution of rein-containing proteins principally in diethylene glycol monoetlwl ether.

Example 3.--A solvent mixture containing 72 parts oi 91% isopropanol and 48 parts of a mixture consisting of 80.9% of propylene glycol and 11.1% of diethylene glycol monoethyl ether was added to 30' parts 01' powdered gluten. The procedure was as in Example l except that the exwas added to 30 parts of powdered gluten. The procedure was as in Example 1 except that the extraction temperature was 140' I". The resulting residue was a solution of the rein-containing proteins principally in the glycol and glycol ether mixture.

At the cessation of boiling as reierred to in the examples some proportion of the lower boiling constituents of the diluent originally used remains in the final solution. The proportion thereof so remaining depends upon the final temperature and the boiling point curve of the mixture. Considerable proportions of the diluent or of its constituents, including water, may remain without causing gelling oi the final solution in reasonable periods of time. This is particularly true where the glycols are used. In this respect the final solutions show difl'erences from solutions oi commercial zein containing large amounts of water as in the prior art, apparently by reason of some changes in the character or structure of the zein-containing proteins of the corn gluten taking place in the process of the present invention, the nature oi which changes is not as yet understood.

In general and influenced by the type and amount of zein protein present, the solutions prepared as described above and the coatingcompositions prepared therefrom have the properties, particularly when heated. oi being able to "give up their solvent or solvents readily, and when applied to a surface they quickly and readily form t-ugh. flexible, nontacky, hard and generally transparent coatings even when retaining some solvent. The coatings formed by these zein-containing solutions and coating compositions are very strong and have adherent properties. In these solutions and coating compositions the film forming properties thereof can be built up to produce films of great toughness,

traction temperature was 180' l". The resulting residue was a solution oi min-containing w teins principally in the glycol and glycol et mixture.

Example L-A solvent mixture containing 60 parts of anhydrous synthetic methanol and 40 parts of a solvent mixture consisting of 75% of diethylene glycol and ct ethylene glycol monoethyl ether was added to parts of nowdered gluten. The procedure was as in Example 1 except that the extraction temperature was 140' F. The resulting residue was a solution of the coin-containing proteins principally in the diethylene glycol and glycol ether mixture.

Emmple 5.A solvent mixture containing 60 parts or a mixture consisting of 80% oi denatured 95% ethyl alcohol ("Pasco) and 20% or water. and parts of a mixture consisting or 75% of diethylene glycol and 25% of ethylene glycol monoethy ether was added to 30 parts of powdered gluten. The procedure was as in Exampir 1 except that the extraction temperature was 140 1''. The resulting residue was a solution of rein-containing proteins principally in the glycol and glycol ether mixture.

Example 0.4 solvent mixture containing Parts of a mixture consisting oi! 90% of 9 isopropanc'. and 10% of water. and 40 parts of a a mixture consisting oi 75% oi diethylene glycol and 25% of ethylene glycol monomethyl ether flexibility. hardness and gloss. Under normal temperature conditions. many of the rein-containing solutions or coating compositions will produce non-blushing coatings on drying, without the aid of supplementary anti-blushing agents as in the prior art.

Th coating compositions prepared from the While my invention has been described in connection with certain specific examples, it is, of course. obvious it is not to be construed as limited to these examples or to the details of the methods set forth therein. since obvious changes in materials, proportions and method details will be apparent from the foregoing.

In the claims, the expression "an elevated temperature in the order of about 1'. to about 170 F." is not to be construed as a precise critical range, but, rather, as a range of temperatures indicative of the elevated temperature at which the extractions may be carried out.

I claim:

1. The method of preparing directly from gluten a solution of min-containing proteins in a solvent or mixture of solvents comprising a glycol ether as an essential solvent constituent for the zein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising a glycol ether and a miscible diluent therefor having a boiling point below about (2., separating the extract from the residual gluten and removing at least part of the diluent from the extract to tom the aloresaid solution.

2. The method oi preparing directly from gluten a solution or rein-containing proteins in a solvent or mixture or solvents comprising a glycol ether as an essential solvent constituent for the rein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order 01' about 120 F. to about 170 F. to extract rein-containing proteins from the gluten, said solvent mixture comprising a glycol ether and a miscible dfluent theretor having a boiling point below about 125 0.. separating the extract from the residual gluten and removing substantially all or the diluent from the extract to form the aforesaid solution.

3. The method oi preparing directly i'rorn gluten a solution 01' stein-containing proteins in a solvent or mixture of solvents comprisin a glycol ether as an essential solvent constituent for the min-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising a glycol ether and a miscible diluent therefor having a boiling point below about 125 C., said diluent comprising a solvent for said rein-containing proteins, separating the extract from the residual gluten and removing at least part oi the diluent from the extract to form the aforesaid solution.

4. The method or preparing directly from gluten a solution or rein-containing proteins in a solvent or mixture of solvents comprising a glycol ether as an essential solvent constituent for the zein-containing proteins. which comprises contacting gluten with a solvent mixture at an elevated temperature in the order oi about 120 1". to about 170 F. to extract min-containing proteins from the gluten, said solvent mixture comprising a glycol ether and a miscible diluent therefor having a boiling point below about 125 (3., said diluent comprising an aqueous solvent for said min-containing proteins, separating the extract irom the residual gluten and removing at least part oi the diluent from the extract to form theaioresaid solution.

5. The method oi preparing directly from sluten a solution or rein-containing proteins in a solvent or mixture of solvents comprising a glycol ether as an essential solvent constituent for the rein-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order of about 120 F. to about 170 F. to extract zein-containing proteins from the gluten, said solvent mixture comprising a glycol ether and a miscible diluent therefor having a boiling point below about 125 0., said diluent being an anhydrous solvent for said zein-containing proteins, separatin: the extract from the residual gluten and removing at least part or the diluent from the extract to form the aforesaid solution.

6. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising diethylene glycol monoethyl ether as an essential solvent constituent for the rein-containing proteins. which comprises contacting gluten with a solvent mixture at an elevated temperature in the order 01' about 120 F. to about 170 F. to extract coin-containing proteins from the gluten, said solvent mixture comprising diethylene glycol monoethyl ether and 01% isopropyl alcohol, separating the extract from the residual gluten and removing at least part of the isopropyl alcohol from the extract to form the aforesaid solution.

7. The method of preparing directly from gluten a solution of zein-containing proteins in a solvent or mixture of solvents comprising diethylene glycol and ethylene glycol monoethyl ether alcohol as essential solvent constituents tor the min-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order 01' about F. to about F. to extract coin-containing proteins from the gluten, said solvent mixture comprising diethylene glycol, ethylene glycol monoethyl ether and methanol, separating the extract from the residual gluten and removing at least part of the methanol from the extract to form the aforesaid solution.

8. The method or preparing directly from gluten a solution of item-containing proteins in a solvent or mixture oi solvents comprising diethylene glycol and ethylene glycol monoethyl ether as essential solvent constituents for the min-containing proteins, which comprises contacting gluten with a solvent mixture at an elevated temperature in the order oi about 120 F. to about 170' 1". to extract rein-containing proteins iron: the gluten, said solvent mixture comprising diethylene glycol, ethylene glycol monoethyl ether, 95% ethyl alcohol and water, separating the extract from the residual gluten andremovingatleastpertoitheethyl alcohol and water from the extract to form the storesaid solution.

BOY E. COLEMAN. 

